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Eukaryotic Cell, June 2004, p. 646-662, Vol. 3, No. 3
1535-9778/04/$08.00+0     DOI: 10.1128/EC.3.3.646-662.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

RasC Plays a Role in Transduction of Temporal Gradient Information in the Cyclic-AMP Wave of Dictyostelium discoideum

Deborah Wessels,1 Rebecca Brincks,1 Spencer Kuhl,1 Vesna Stepanovic,1 Karla J. Daniels,1 Gerald Weeks,2 Chinten J. Lim,2 George Spiegelman,2 Danny Fuller,3 Negin Iranfar,3 William F. Loomis,3 and David R. Soll1*

W. M. Keck Dynamic Image Analysis Facility, Department of Biological Sciences, The University of Iowa, Iowa City, Iowa 52242,1 Department of Microbiology and Immunology, The University of British Columbia, Vancouver, British Columbia,2 Department of Biology, University of California, San Diego, La Jolla, California 920393

Received 8 October 2003/ Accepted 8 January 2004

To define the role that RasC plays in motility and chemotaxis, the behavior of a rasC null mutant, rasC–, in buffer and in response to the individual spatial, temporal, and concentration components of a natural cyclic AMP (cAMP) wave was analyzed by using computer-assisted two-dimensional and three-dimensional motion analysis systems. These quantitative studies revealed that rasC– cells translocate at the same velocity and exhibit chemotaxis up spatial gradients of cAMP with the same efficiency as control cells. However, rasC– cells exhibit defects in maintaining anterior-posterior polarity along the substratum and a single anterior pseudopod when translocating in buffer in the absence of an attractant. rasC– cells also exhibit defects in their responses to both the increasing and decreasing temporal gradients of cAMP in the front and the back of a wave. These defects result in the inability of rasC– cells to exhibit chemotaxis in a natural wave of cAMP. The inability to respond normally to temporal gradients of cAMP results in defects in the organization of the cytoskeleton, most notably in the failure of both F actin and myosin II to exit the cortex in response to the decreasing temporal gradient of cAMP in the back of the wave. While the behavioral defect in the front of the wave is similar to that of the myoA–/myoF– myosin I double mutant, the behavioral and cytoskeletal defects in the back of the wave are similar to those of the S13A myosin II regulatory light-chain phosphorylation mutant. Expression array data support the premise that the behavioral defects exhibited by the rasC– mutant are the immediate result of the absence of RasC function.

* Corresponding author. Mailing address: 302 BBE, Department of Biological Sciences, The University of Iowa, Iowa City, IA 52242. Phone: (319) 335-1117. Fax: (319) 335-2772. E-mail: david-soll{at}uiowa.edu.

Eukaryotic Cell, June 2004, p. 646-662, Vol. 3, No. 3
1535-9778/04/$08.00+0     DOI: 10.1128/EC.3.3.646-662.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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